Multiple digital PCR chip and use method thereof

Abstract

The invention relates to a multiple digital PCR chip and a use method thereof. The chip does not need to be driven by a precise micro-pump and be controlled by a complex micro-valve and does not need a complex macro-micro interface. The chip realizes simple, fast, low cost and large-scale autodecomposition and quantitative uniform distribution of samples. Compared with the existing commercial PCR chip, the chip greatly reduces the complexity and operating cost of the digital PCR system, simplifies the experimental operation and is free of professional training of an operator. The chip realizes the independence of each PCR reaction unit through microcavity arrays, is free of a surfactant and prevents the influence caused by an additive on the reaction system. Through integration of the isolated microcavity array zones and pre-deposition of primers corresponding to different specific target molecules in the microcavities in different zones, the chip can realize multiple detection through one step. The chip has the advantages of operation simpleness, low cost and strong multiple detection capacity and can promote development and wide application of the digital PCR technology.

Description

technical field [0001] The invention belongs to the technical field of microfluidic chip analysis, and in particular relates to a multiple digital PCR chip and a using method thereof. Background technique [0002] Polymerase chain reaction (PCR) is an in vitro nucleic acid amplification technology, which can efficiently and rapidly amplify trace target nucleic acids in vitro for detection and application. With the continuous development of molecular biology technology, PCR technology is constantly updated, and is currently transforming from traditional quantitative PCR to digital PCR. The principle of digital PCR is to decompose the sample into thousands or even millions of droplets (each droplet acts as an independent PCR reaction unit), until each droplet contains one or zero target molecules , when all droplets undergo PCR amplification at the same time, the droplets containing the target molecule will generate a positive signal due to the amplification of the target mol...

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Problems solved by technology

Zhong et al. used the same fluorescein FAM to label probes of different mutation types, and then added them to the PCR mixture at different concentrations, and used the fluorescence intensity of the positive milk droplets after the digital PCR reaction to determine which mutations occurred and the types of mutations. [Zhong Q, Bhattacharya S, Kotsopoulos S, et al.Multiplex digital PCR: breaking the onetarget per color barrier of quantitative PCR.Lab on a Chip,2011,11(13):2167-2174.], but due to PCR amplification The fluorescence intensity of the product is often easily interfered by other factors, so this method has certain shortcomings in the accuracy and reliability of multiplex digital PCR detection

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